BIOFUNCTIONAL EVALUATION OF A HYDROGEN BOND STABILIZING THE β-TURN IN THE ACYCLIC PART OF OXYTOCIN
Corresponding Author
J. ROY
Department of Physiology and Biophysics and the Center for Polypeptide and Membrane Research, Mount Sinai Medical and Graduate Schools of the City University of New York, New York, U.S.A.
Department of Physiology and Biophysics Mount Sinai Medical and Graduate Schools of the City University of New York New York, NY 10029 U.S.A.Search for more papers by this authorM. JOHNSON
Department of Physiology and Biophysics and the Center for Polypeptide and Membrane Research, Mount Sinai Medical and Graduate Schools of the City University of New York, New York, U.S.A.
Search for more papers by this authorD. GAZIS
Department of Physiology and Biophysics and the Center for Polypeptide and Membrane Research, Mount Sinai Medical and Graduate Schools of the City University of New York, New York, U.S.A.
Search for more papers by this authorI.L. SCHWARTZ
Department of Physiology and Biophysics and the Center for Polypeptide and Membrane Research, Mount Sinai Medical and Graduate Schools of the City University of New York, New York, U.S.A.
Search for more papers by this authorCorresponding Author
J. ROY
Department of Physiology and Biophysics and the Center for Polypeptide and Membrane Research, Mount Sinai Medical and Graduate Schools of the City University of New York, New York, U.S.A.
Department of Physiology and Biophysics Mount Sinai Medical and Graduate Schools of the City University of New York New York, NY 10029 U.S.A.Search for more papers by this authorM. JOHNSON
Department of Physiology and Biophysics and the Center for Polypeptide and Membrane Research, Mount Sinai Medical and Graduate Schools of the City University of New York, New York, U.S.A.
Search for more papers by this authorD. GAZIS
Department of Physiology and Biophysics and the Center for Polypeptide and Membrane Research, Mount Sinai Medical and Graduate Schools of the City University of New York, New York, U.S.A.
Search for more papers by this authorI.L. SCHWARTZ
Department of Physiology and Biophysics and the Center for Polypeptide and Membrane Research, Mount Sinai Medical and Graduate Schools of the City University of New York, New York, U.S.A.
Search for more papers by this authorAbstract
In a continued effort to determine the importance of the hydrogen bonds for stabilization of the biologically active conformation of oxytocin, deamino-[9-glycolicamide] oxytocin was synthesized in order to study, in this respect, the hydrogen bond between the peptide N-H of Gly9 and the C=O of Cys6. In this analog the amide linkage between residues at positions 8 and 9 is replaced by an ester. Thus, the residue at position 9 cannot be involved in hydrogen bond formation with the C=O of Cys6. Deamino-[9-glycolicamide] oxytocin exhibited 134 ± 13 U/mg and 355 ± 48 U/mg of uterotonic activity in absence and in presence, respectively, of Mg2+, 108 ± 8 U/mg of milk-ejecting activity, 0.35 ± 0.03 U/mg of pressor activity and 2.5 ± 0.1 U/mg of antidiuretic activity. It is concluded that the hydrogen bond under question is not critical for the conformation required for biofunctional interaction of oxytocin with its receptors in the uterus, mammary gland and other target organs.
REFERENCES
- Ballardin, A., Fischman, A.J., Gibbons, W.A., Roy, J., Schwartz, I.L., Smith, C.W., Walter, R. & Wyssbrod, H.R. (1978) Biochemistry 17, 4443–4454.
- Bisset, G.W., Clark, B.J., Haldar, J., Harris, M.C., Lewis, G.P. & Rocha e, Silva M., Jr. (1967) Brit. J. Pharm. Chemother. 31, 537–549.
- Bradbury, A.F., Burgen, A.S.V., Feeney, J., Roberts, G.C.K. & Smyth, D.G. (1974) FEBS Lett. 42, 179–182.
- Brewster, A.I.R. & Hruby, V.J. (1973) Proc. Natl. Acad. Sci. US 70, 3806–3809.
- Brewster, A.I.R., Hruby, V.J., Spatola, A.F. & Bovey, F.A. (1912a) Biochemistry 12, 1643–1649.
- Brewster, A.I.R., Hruby, V.J., Glasel, J.A. & Tonelli, A.E. (1973b) Biochemistry 12, 5294–5304.
- Cash, W.D., Mahaffey, L.McC., Buck, A.S., Nettleton, Jr., D.E., Romas, C. & du Vigneaud, V. (1962) J. Med. Pharm. Chem. 5, 413–423.
- Chan, W.Y. & Kelley, N. (1967) J. Pharmacol. Exp. Ther. 156, 150–158.
- Dekanski, J. (1952) Brit. J. Pharmacol. 7, 567–572.
- Feeney, J., Roberts, G.C.K., Rockey, J.H. & Burgen, A.S.V. (1971) Nature New Biol. 232, 108–110.
- Ferrier, B.M., Jarvis, D. & du Vigneaud, V. (1965) J. Biol. Chem. 240, 4264–4266.
- Glickson, J.D., Urry, D.W. & Walter, R. (1972) Proc. Natl. Acad. Sci. US 69, 2566–2569.
- Glickson, J.D., Rowan, R., Pitner, T.P., Dadok, J., Bothner-By, A.A. & Walter, R. (1976) Biochemistry 15, 1111–1119.
- Griffin, J.H., Alazard, R., Dibello, C., Sala, E., Mermet-Bouver, R. & Cohen, P. (1975) FEBS Lett. 50, 168–171.
- Holton, P. (1948) Brit. J. Pharmacol. 3, 328–334.
- Hruby, V.J., Deb, K.K., Spatola, A.F., Upson, D.A. & Yamamoto, D. (1979) J. Am. Chem. Soc. 101, 202–212.
- Jeffers, W.A., Livesey, M.M. & Austen, J.H. (1942) Proc. Soc. Exp. Biol. Med. 50, 184–188.
- Johnson, L.F., Schwartz, I.L. & Walter, R. (1969) Proc. Natl. Acad. Sci. US 64, 1269–1275.
- Live, D.H., Wyssbrod, H.R., Fischman, A.J., Agosta, W.C., Bradley, C.H. & Cowburn, D. (1979) J. Am. Chem. Soc. 101, 474–479.
- Lowry, O.H., Rosebrough, N.J., Farr, A.L. & Randall, R.J. (1951) J. Biol. Chem. 193, 265–275.
- Meraldi, J.-P., Hruby, V.J. & Brewster, A.I.R. (1977) Proc. Natl. Acad. Sci. US 74, 1373–1377.
- Mühlemann, M., Titov, M.I., Schwyzer, R. & Rudinger, J. (1972) Helv. Chim. Acta. 55, 2854–2860.
- Munsick, R.A. (1960) Endocrinology 66, 451–457.
- Porath, J. & Flodin, P. (1959) Nature 183, 1657–1659.
- Roy, J., Roy, U., Gazis, D.C. & Schwartz, I.L. (1979) Proc. Natl. Acad. Sci. US 76, 3309–3313.
- Sawyer, W.H. (1958) Endocrinology 63, 694–698.
- Schild, H.O. (1942) J. Physiol. (London) 101, 115–130.
- Schwartz, I.L. (1974) in The Chemistry of Polypeptides ( P.G. Katsoyannis, ed.), Chapter 14, pp. 297–313, Plenum, New York .
- Shemyakin, M.M., Ovchinnikov, Y.A., Ivanov, V.T. & Kiryushkin, A.A. (1963) Tetrahedron 19, 995–1000.
- Spackman, D.H., Stein, W.H. & Moore, S. (1958) Anal. Chem. 30, 1190–1206.
- Stewart, F.H.C. (1965) Aust. J. Chem. 18, 1089–1094.
- Urry, D.W. & Walter, R. (1971) Proc. Natl. Acad. Sci. US 68, 956–958.
- Urry, D.W., Ohnishi, M. & Walter, R. (1970) Proc. Natl. Acad. Sci. US 66, 111–116.
- van Dyke, H.B., Sawyer, W.H. & Overweg, N.I.A. (1963) Endocrinology 73, 637–639.
- Walter, R. (1977) Fed. Proc. 36, 1872–1878.
- Walter, R., Schwartz, I.L., Darnell, J.H. & Urry, D.W. (1971) Proc. Natl. Acad. Sci. US 68, 1355–1359.
- Walter, R., Prasad, K.U.M., Deslauriers, R. & Smith, I.C.P. (1973) Proc. Natl. Acad. Sci. US 70, 2086–2090.
- Walter, R., Smith, I.C.P. & Deslauriers, R. (1974) Biochem. Biophys. Res. Commun. 58, 216–221.
- Walter, R., Wyssbrod, H.R. & Glickson, J.D. (1977) J. Am. Chem. Soc. 99, 7326–7332.
- Wyssbrod, H.R., Ballardin, A., Schwartz, I.L., Walter, R., van Binst, G., Gibbons, W.A., Agosta, W.C., Field, F.H. & Cowburn, D. (1977) J. Am. Chem. Soc. 99, 5273–5276.
- Yamashiro, D. (1964) Nature 201, 76–77.
- Yamashiro, D., Gillessen, D. & du Vigneaud, V. (1966) J. Am. Chem. Soc. 88, 1310–1313.
- Zahn, H. & Rexroth, E. (1955) Z. Anal. Chem. 148, 181–186.
